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A family 5 β-mannanase from the thermophilic fungus Thielavia arenaria XZ7 with typical thermophilic enzyme features

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Abstract

A novel β-mannanase gene, man5XZ7, was cloned from thermophilic fungus Thielavia arenaria XZ7, and successfully expressed in Pichia pastoris. The gene (1,110 bp) encodes a 369-amino acid polypeptide with a molecular mass of approximately 40.8 kDa. The deduced sequence of Man5XZ7 consists of a putative 17-residue signal peptide and a catalytic module belonging to glycoside hydrolase (GH) family 5, and displays 76 % identity with the experimentally verified GH 5 endo-β-1,4-mannanase from Podospora anserina. Recombinant Man5XZ7 was optimally active at 75 °C and pH 5.0 and exhibited high activity at a wide temperature range (>50.0 % activity at 50–85 °C). Moreover, it had good adaptability to acidic to basic pH (>74.1 % activity at pH 4.0–7.0 and 25.6 % even at pH 9.0) and good stability from pH 3.0 to 10.0. These enzymatic properties showed that Man5XZ7 was a new thermophilic and alkali-tolerant β-mannanase. Further amino acid composition analysis indicated that Man5XZ7 has several characteristic features of thermophilic enzymes.

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Acknowledgments

This research was supported by the National Science Foundation for Distinguished Young Scholars of China (31225026) and the National Natural Science Foundation of China (31172235) and the China Modern Agriculture Research System (CARS-42).

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Authors and Affiliations

  1. Key Laboratory for Feed Biotechnology of the Ministry of Agriculture, Feed Research Institute, Chinese Academy of Agricultural Sciences, No.12 Zhongguancun South Street, Beijing, 100081, People’s Republic of China

    Haiqiang Lu, Pengjun Shi, Huiying Luo, Yaru Wang, Peilong Yang & Bin Yao

  2. College of Biotechnology, Tianjin University of Science and Technology, Tianjin, 3000457, People’s Republic of China

    Huitu Zhang

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  1. Haiqiang Lu
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  2. Huitu Zhang
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  3. Pengjun Shi
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  4. Huiying Luo
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  5. Yaru Wang
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  6. Peilong Yang
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  7. Bin Yao
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Correspondence to Huitu Zhang or Bin Yao.

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Lu, H., Zhang, H., Shi, P. et al. A family 5 β-mannanase from the thermophilic fungus Thielavia arenaria XZ7 with typical thermophilic enzyme features. Appl Microbiol Biotechnol 97, 8121–8128 (2013). https://doi.org/10.1007/s00253-012-4656-1

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  • DOI: https://doi.org/10.1007/s00253-012-4656-1

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